CC BY
9Our preliminary studies have shown that drug-containing CSF with KXS in rats can improve the PC12 cell injury significantly induced by serum-free culture and promote cell survival adherent.Which indicates KXS was nutritious and protective to PC12 cells. These results suggest that the drug-containing CSF with KXS may exist active substances with neural growth factor-like effects[7]. Therefore,this also shows that the material basis of its efficacy exists inevitable objectively. The basic problem to be solved is that whether the active substance (group) is the prototype of the product or new metabolic substance of KXS and how about it's basic information.
As the content has been low when the drug absorbed into the bloodstream,coupled with the special structure of the Blood Brain Barrier makes it difficult to through drugs,which leading drug content in cerebrospinal fluid of animals is minimal and causing lower response peaks. We found three iconic migration component in the drug-containing CSF that had gavaged KXS.We also found thatthe three compounds were both derived from the prototype components of Ginseng in KXS according to its chromatographic characteristics and contrast to composition in vitro. These three compounds may be substance that have more direct neuroprotective effect,and at present,the structural and activity identification of them is being developedactively. References
1.Bian Huimin,Guo Haiying,Huang Yufang,et al. Effect of Kaixin-san on learning and memory of mice in four animal models.Chinese Journal of Experimental Traditional Medical Formulae,1999,5(5):51-53.
2.Mai weili, Wang Qiong, Sun lihua, et al.Effect of Kaixinsan on learning and memory of mice after sleep deprivation. LiSHiZhen-Medicine And Materta Research. 2010,22(10):2331-2333.
3.Huang yufang, Bian huimin, Gong jiening,et al. Effect of Kaixingsan on SOD and MDA contents in four animal models. Journal Of NanJing University Of TCM, 1999, 15 (3) : 151.
4 Huang fang,Huang zijie,Wang jiali.The study of Anti-lipid peroxidation of each component of KXS in vitro [J]. Chinese Journal of Experimental Traditional Medical Formulae, 2007, 13 (1)55-57.
5 Zeng yidan,Huang fang,Wang jiali. The modern pharmacological research of KXS [J]. Strait Pharmaceutical Journal, 2006, 18(5):12-14.
6 Wen wei,Zhang chao,Hang shuming,et al.The improvement of KXS Containing serum on Ap-induced injury in PC12 cells [J]. Information on Traditional Chinese Medicine, 2012, 29(4):80-81.
7 Li fudong,Wang yanhua,Huang shuming,et al. The growth-promoting effect of KXS drug-containing cerebrospinal fluid on serum-free culture PC12 cell [J]. Journal of Traditional Chinese Medicine, 2012, 40(6):12-14.
Studies on Amide Alkaloids of theViscum coloratum (Kom.) Nakai f.
Rubroaurantiacum Kitag
Yong-hui Sun1, Ying-li Ma 2, Yong Ling 2
(1.Research Institute of Chinese medicine, Heilongjiang university of Chinese medicine; 2.Dept of TCM, Heilongjiang university of Chinese medicine, harbin 150040,China )
Abstract: Using chromatography technique two constituents were isolated from Viscum coloratum (Kom.) Nakaif. Rubroaurantiacum Kitag for the first time. On the basis of chemical methods and spectroscopic analysis their structure were characterized as N-butyl amido cinnamyl amine and N-cinnamoyl spermidine (new compound) .
Key words: Viscum coloratum (Kom.) Nakai f. Rubroauran tiacum Kitag ; amide alkaloids; N-butyl amido cinnamyl amine; N-cinnamoyl spermidine
1 Introduction
Viscum coloratum (Kom.)Nakai, which belongs to genus Viscum of the Loranthaceae Family, is distributed widely across China, especially in Northern China. It have the effects of dispelling pathogenic wind and dampness, nourishing liver and kidneys, strengthening tendons and bones, as well as nourishing blood for preventing miscarriage according to the theory of TCM.There are around 30 species of genus Viscum, in which llspeciesare distributed in most provinces of China.The host plant includes elm, birch, bass, maple, poplar and willow etc. There are two variety species in northern area of China. One is V. coloratum (Kom.) Nakai f. Rubroauran tiacum Kitag with red fruits; another is V. coloratum (Kom.) Nakai f. lutescens Kitag with yellow fruits. Various active components are found in mistletoe, most of which are flavonoids, alkaloids and macromolecule compounds. Besides, triterpenoids, organic acids, volatile oils, several phenylpropanoidsandsterolsarealsoisolatedfrommistletoeextracts. According to the modern Pharmacological research, the gradients of alkaloids show medicinal effects of anti-tumor. In order to find the Pharmacological-active compounds of mistletoe, exploit and utilize this medicinal plant resource, the studies on alkaloids constituents of V. coloratum (Kom.) Nakai f. Rubroauran tiacum Kitag growing in Baoqing county, HeilongjiangProvince were carried out.
2 Experimental
2.1 Experimentalmaterial
2.1.1 Plant material
V. coloratum (Kom.) Nakai f. Rubroauran tiacum Kitag plants were collected from mountains in Baoqing county, HeilongjiangProvince.
2.1.2 Instruments
BRUKER 600 Ultra Shield™ NMR spectrometer; Bruker micrOTOF-Q 125 mass spectrometer.
2.2 Extraction and Isolation
The particle of dried stems and leaves of V. coloratum (Kom.) Nakai f. R. (20kg) were extracted three times with 75%EtOH (3 X 60L) by permeated methods. After concentrating under reduced pressure ,the extracts was dissolved with 2%HCl/H2O.After filtrated, NH3 ■ H2O was used to adjust the acid water extracts pH8~9.Then, the alkaline extracts solution were extracted with CHCl3. Solvent was removed to give CHCl3 extracts(70g).Compound I and N were obtained by repeatedly column chromatography over silica gel and ODS C18.
3 Results and Discussion
Compound I was obtained as colorless flake crystal. Dragendorff reaction was positive.Its molecular formula was assigned as C13H18N2O on the basis of the ESI-MS showed an M+ peak at m/z 219[M+H]+ ,which was confirmed by 1H-NM Rand 13C-NMR.
The 1H-NMR spectrum of I showed the presence of single-substituted benzene ring at 57.53(2H, m, H-2, 6),7.35(3H, m, H-3* 4, 5). The signal at 57.50(1H, d, J=16.2,H-7)and 6.57(1H, d, J=16.2, H-8)confirmed the existence of olefinic bond proton in trans-form. The signal at S3.31(2H, t, J=3.6Hz, H-1') and 2.76 (2H, t, J=7.2, H-4')suggested the presence of nitrogen methylene.There are four proton signals of methylene at S1.59(4H, m, H-2\ 3').
13
In the C-NMR spectrum, the signal of S168.7(C-1) suggested carbonyl of amide. The signal 8141.7(C-7) and 121.8(C-8)showed olefinic bond carbon. At 5136.3(C-1),130.8(C-4),129.9(C-2s 6) and 128.7 (C-3* 5) are the signals of benzene ring carbon. There are two signals of nitrogen methylene carbon at S4L5(C-1'), 40.0 (C-4') , and two signals of methylene carbon at 29.2(C-2') and 27.7(C-3'). (structure of compound I see Fig. 1)
Fig. 1 structure of compound I Fig. 2 structure of compound II
Compound N was obtained as colorless flake crystal. Dragendorffreaction was positive. Its molecular formula was assigned as C16H25N3O on the basis of the ESI-MS showed an M+ peak at m/z 276[M+H]+ ,which was confirmed by 1H-NMR and 13C-NMR. 1H-NMR, 13C-NMR spectrum of compound N is similar to compound I in low field which proved the presence of cinnamoyl. Seven methylene carbon appeared in high field of DEPT spectrum. Among them there are four methylene carbon signalwhich linked with nitrogen and three methylene carbon signal. 1H-NMR spectrum revealed the existence of amino proton at 52.88 * 2.71 and amide proton at 57.39 which
13
superposed with benzene ring proton. H-NMR* C-NMR* DEPT and HMBC spectrum data of compound 11 see table 1. (structure of compound 11 see Fig. 2 and HMBC correlations see Fig. 3)
Fig. 3 HMBC correlations of compound N Tab 1NMR data of compound II
No. 13C-NMR 1 H-NMR DEPT HMBC
1 7.39
2 37.0 3.46 (2H, t, J=6Hz) CH2 H-3v H-4
3 27.7 1.99 (2H, m) CH2 H-2v H-4
4 46.4 3.07 (2H, m) CH2 H-2v H-3v H-6
5 2.88 (1H, s)
6 48.2 3.07 (2H, m) CH2 H-4v H-7v H-8
7 25.6 1.81 (2H, m) CH2 H-6v H-8v H-9
8 24.3 1.81 (2H, m) CH2 H-6v H-7v H-9
9 40.0 3.01 (2H, t, J=7.2Hz) CH2 H-7v H-8
10 2.71 (2H, s)
1' 136.0 C H-8' v H-3'v H-5'
2' 130.0 7.57 (1H, m) CH H-3' v H-4'v H-6'
3' 128.9 7.39 (1H, m) CH H-2' v H-5'v H-4'
4' 131.1 7.39 (1H, m) CH H-2' v H-6'
5' 128.9 7.39 (1H, m) CH H-3' v H-4'v H-6'
6' 130.0 7.57 (1H, m) CH H-2' v H-5'v H-4'
7' 142.3 7.57 (1H, ) CH H-8' v H-2'v H-6'
8' 121.2 6.66 (1H, d, J=15.6) CH H-7'
9' 169.5
C
H-7'v H-8'v H-2
References:
[1]Delectis Florae Reipublicae Popularis Sinicae, Agendae Academiae Sinicae Edita.Flora Reipublicae Popularis Sinicae, Tomus 24. Beijing: Science Publishing House,1986, 147-148.
[2]Wu J R. Research on the Original Plants of Crude Drug"Sang Ji Sheng"(Ramulus Taxilli).Chinese Traditional Herbal Drugs, —1981—12(5):32
[3]Peng H Y, Zhang Y H, Han Y et al. Studies on the anticancer effects of total alkaloid fromViscum coloratum. China Journal of Chinese Materia Medica. —2005—30(5):381~387.
Two NewGlycosides from the Fruits ofMorinda eitrifolia L.
Hong-CaiZhang1^, Yu Wang1, Shu-Min Liu 1*
*Academy of traditional Chinese medicine,Heilongjiang University of Chinese Medicine, Harbin 150040, China; E-Mails: [email protected] (H.-C.Z.); [email protected] (Y.W.)
*Authors to whom correspondence should be addressed; E-Mails: [email protected] (S.-M.L.);[email protected] (L.L.); Tel.: +86-139-451-33028 (S.-M.L.).
Abstract: To study the chemical constituents of the fruits of noni(Morinda citrifoliaL.), and find novelcompounds, an n-butanol extract of the ethanol soluble fraction was subjected to repeated silica gel and ODS column chromatography and HPLC. Two new glycosides were isolated and their structures elucidated by NMR and HRFAB-MS spectrometry as(2£',4£',7Z)-deca-2,4,7-trienoate-2-O-ß-D-glucopyranosyl-ß-D-glucopyra-noside (1) andamyl-1-O-ß-D-apio-furanosyl-1,6-O-ß-D-glucopyranoside (2), respectively.
Keywords:Morinda citrifoliaL. ;glycosides
1. Introduction
Morinda citrifolia L. (family Rubiaceae), also known as Tropical Radix Morindae Officinalis (RMO) and Medicinal Morinda Root Seasonal Fruit, is usually a small tree or bush occurring in the South Pacific tropical islands and widely distributed in the Hainan Province and Paracel Islands of China and in Taiwan. The fruits of Morinda citrifolia L. are oval and juicy with a strong odor, and have been used for a long time as a medicinal plant in Southeast Asia and the Pacific Islands. All parts of the plant can been used, including fruit, leaf, root, bark, flower, stem and seed[1]. Reported traditional uses include as a treatment of boils, abscesses, and inflammations of various origins, fungal infections, and constipation as well as diarrhea[2,3]. Pharmacological research has revealed a number of biological activities in recent years, such as anticancer[4], anti-inflammation[5], antioxidant[6], liver protection[7], and anti-AIDSproperties [8]. In this work silica gel column chromatography was employed to separate the glucoside constituents of an n-butanol extract of the ethanol soluble fraction of Morinda citrifolia L. fruits. 2D-NMR techniques, HR-ESI-MS and hydrolytic reactions were used to elucidate the structures of the extracted compounds.
2. Results and Discussion
Compound 1 (Figure 1) was obtained as a white powder (15.6 mg). The molecular formula was determined to be C22^4OBby the HR-FAB-MS [M+H]+ peak at m/z507.2071. Acid hydrolysis
1 13
of 1 only gave D-glucose. The H and C-NMR spectra of 1 indicated an alkenoic acid ester moiety and two glucose groups. The alkenoic acid ester moiety was confirmed by 1H-NMR (Table 1) signals at 5h6.00 (1H,d, J = 15.2Hz), 7.74(1H,ddd, J = 15.2,11.6,0.8Hz), 6.21 (1H, t, J = 11.3 Hz),
